Table 2.
Percent APCs containing BCG-GFP by flow cytometry | Percent affinity-sorted cells containing BCG-GFP by flow cytometry | CFU/affinity-sorted cells | Detection efficiency | |
---|---|---|---|---|
Formula*2 | mean (R6/(R2 + R3 + R4)) | mean (R6′/R1) | mean (CFU/R1) | mean (R6′/CFU) |
CD11b affinity-sorted cells | 0.60 +/− 0.21% | 0.29 +/− 0.18% | 0.9 +/− 0.56% | 38 +/− 0.17% |
CD11c affinity-sorted cells | 0.52 +/− 0.26% | 0.35 +/− 0.22% | 1.1 +/− 0.47% | 42 +/− 0.37% |
Data are shown as mean +/− SD. Data are from 3 separate experiments with 2–5 mice per group (pooled to generate CD11b or CD11c affinity-sorted samples).
R6 events in column 2 are BCG-GFP+ events derived from R2 + R3 + R4, and the proportion of lung APCs that were BCG-GFP+ was determined as R6/(R2 + R3 + R4). For CFU determination, samples were taken from total CD11b or CD11c affinity-sorted cell preparations and included cells both inside and outside R2 + R3 + R4 (strictly represented by R0 events and effectively represented by the R1 gate that excludes only subcellular debris), necessitating comparison of CFU to BCG-GFP+ events derived from R1 (not just R2 + R3 + R4). Therefore, R6′ events in columns 3 and 5 are BCG-GFP+ events derived from R1. CFU values were determined by culture of affinity-sorted cells. R1 was determined by flow cytometry analysis in column 3 and by cell count with a hemacytometer in column 4. Detection efficiency was calculated as mean (R6′/CFU) as opposed to (mean R6′/R1)/(mean CFU/R1).